BACKGROUND: Donor complications have been detected following autologous tendon transplantation for posterior cruciate ligament reconstruction. Although artificial tendon development and tissue-engineered tendon have achieved great progresses, there are some issues in clinical application. Since 1980's, allogenic tendon transplantation has aroused increasing attention. OBJECTIVE: To explore the selection of allogenic tendon materials and the effect of their application on reconstructing posterior cruciate ligament. METHODS: A total of 17 patients with posterior cruciate ligament injury of knee joint were treated with cryopreserved allogenic tendon by Tibial-inlay technique. During the operation, two tracts of tendons soaked in gentamicin saline for 15 minutes were conduplicated, and one end of the tendon was cancellous bone screw and fixed to the tibia attachment point of posterior cruciate ligament, and the other end was introduced into the joint through retention suture. The posterior joint capsule was repaired. The patient was placed at supine position, and the knee was flexed for 90°. The other end of the graft was introduced to femoral tunnel, and anterior drawer was tensed, and fixed by screw. RESULTS AND CONCLUSlN: The preoperative posterior drawer test of patients was >2+, including 7 cases of 3+ and 6 of 4+. The postoperative posterior drawer test was 0 in 4 cases, 1+ in 8 cases, 2+ in 4 cases and 3+ in 1 case, suggesting the posterior movement of the knee joint was significantly improved. Lysholm scores of patients were (48.5±4.3) points before operation and (88.3±5.4) points after operation. Results show that cryopreserved allogenic tendon by Tibial-inlay technique could restored function of posterior cruciate ligament with a favorable effect.

We here systematically summarize our practice and its effectiveness in the creation and development of the discipline proteomics in Academy of Military Medical Sciences,and the establishment of a system for identifying and training talented researchers in proteomics.The methods we used to train the researchers and to develop a vigorous team are discussed.These practices contributed much to the development of proteomics in China.

Objective To investigate therapeutic effects of phlorizin on the early stage of diabetic retinopathy(DR)in diabetic db/db mice.Methods The 24 db/db male mice were divided into 3 groups of phlorizin-treated diabetic group(20mg/kg of phlorizin by intragastric gavage)(DMT,n =8),diabetic group(normal saline solution by intragastric gavage)(DM,n=8),with C57BLKS/J db/ m mice as the control group(CC,n=8)for ten weeks.After ten weeks,all mice were sacrificed and the retina was isolated.The specimen was embedded in paraffin wax and sliced.The sections were dyed with Sudan red and observed for retinal morphological change.Meanwhile,the expressions of GSK-3β and phospho-GSK-3β in retinas were determined by western blot.Results During the study,the CC group showed good condition.The DM group featured with polydipsia,polyphagia and a rapid increase of body weight.Phlorizin-treated diabetic mice displayed much better than the DM group.Compared with CC group,DM group showed cells proliferation in capillary endothelia and obvious thickening in basement membrane.After administration of phlorizin,the vascular lesion of central regions was remarkably improved.The expression level of phospho-GSK-3β protein was lower in DM group than in control group(76.2±15.8 vs.255.4 ± 10.7,t =16.30,P＜0.01)and was higher in Phlorizin-treated DM group(188.5±18.4)than in DM group(76.2± 15.8,t=8.05,P＜0.01).Conclusions Phlorizin decreases phospho-GSK-3β3expression in retina in db/db diabetic mice and suppresses the GSK-3βactivity,which leads to the protection of DR.

AIM: To facilitate the suicide gene delivery into neoplasm, a chimeric gene of HSV-tk and green fluorescent protein (gfp) was constructed. METHODS: Molecular cloning technique was used to construct this kind of eukaryotic vector. The internal ribosome entry site (IRES) of encephalomyocarditis virus (EMCV), which could coordinate expression of two genes in a single vector, was optioned. By using liposome-mediated transfection, eukaryotic expression vector tgCMV/hytk-IRES-gfp was transfected into human bladder carcinoma cells EJ. RESULTS: A bicistronic eukaryotic vector carrying gfp and hygromycin phosphotransferase-thymidine kinase fusion (hytk) gene was constructed. The results of PCR and microscopy detection show that the hytk-IRES-gfp gene was successfully transferred into EJ cells. There were no differences in the growth pattern or the morphology between EJ and EJ/hytk-GFP cells. In vitro experiments demonstrated dose- and time-dependent cell killing by transduction of the hytk-IRES-gfp gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) was 2.16 mg/L in treatment with GCV for 72 hours. CONCLUSION: These results suggest that this new kind of eukaryotic vector could serves as a new tool and method for neoplasm gene therapy.

Background Ultraviolet radiation is one of factors of the formation of age-related cataract.Indole-3-carbinol(I3C) is a plant chemical material with inhibitory effect on oxidative-induced cell damage and formation of amyloid fibrils,and the oxidative damage and amyloid fibrils are associated with cataract.However,the relationship between I3C and α-crystallin is in study. Objective This study was to evaluate the effects of ultraviolet-B laser irradiation on the secondary structure of α-crystallin and to explore the protection of I3C to chaperone activity of α-crystallin. Methods The fresh eyeballs were obtained from 1-year-old cattle to prepare the purified lens α-crystallin by gel chromatography.α-Crystallin was isolated from cattle lenses using gel chromatography.The purified α-crystallin was collected using fast protein liquid chromatography ( FPLC ) and exposed to 1:308 nmultraviolet-B at different irradiation intensities ( 23.75,118.75,475.00,1187.50,2375.00,4750.00,11 875.00,23 750.00 mJ/cm2 ) and then to ultraviolet-B 2:308 nm with irradiation intensities of 28 535.00,6730.00,3435.00,1910.00,1040.00 mJ/cm2.Ultraviolet-absorbance spectra,tryptophan fluorescence and N-formylkynurenine (N-FK)fluorescence spectra of both irradiated and non-irradiated α-crystallin were measured.I3C at the concentrations of 50 μmol/L and 100 μmoL/L were added to the α-crystallin solution to perform a catalase (CAT) thermal aggregation to confirm the chaperone activity of the α-crystallin,and the α-crystallin solution without any I3C was used as control.The ratios of A360 between various intervene groups with control group were calculated using spectrophotometry.Results The A280 values of the α-crystallin declined to 10％ at the ultraviolet-B irradiation intensity of 1187.5 mJ/cm2 and that at the intensity of 23.75 J/cm2 lowed to 2％.A negative correlation was seen between the ultraviolet-B irradiation intensity and the A280 value of the α-crystallin (R2=0.925 ) and a positive correlation was found between ultraviolet-B with N-FK ( R2 =0.949 ).Ultraviolet-B irradiation intensity showed a negative correlation with Trp fluorescence intensity (R2 =0.996 ).CAT hot condensed experiment revealed that after addition of different concentrations of indole-3-carbinol,the relative A360 values at various ultraviolet-B irradiation group were significantly higher than those of the control group (P =0.000),and the decreasing degree of chaperone activity of α-crystallin was lower than that of the control group ( P =0.000 ). Conclusions The study suggests that I3C can protect the chaperone activity of α-crystallin from photooxidation,and the ultraviolet-B laser may be a good exposure source compared with ultraviolet lamp.The ultraviolet-B laser irradiation causes the alteration of structure and chaperone activity of α-crystallin.

Objective To explore the preoperative localization diagnosis and surgical techniques of intractable occipital lobe epilepsy.Methods Retrospectively studied 37 patients diagnosed as occipital lobe epilepsy and underwent focal occipital resections for epilepsy.The semiology,scalp electroencephalography,MRI,fluorodeoxyglucose-positron emission tomography(FDG-PET),and intracranial EEG monitoring were used to localize the epileptogenic zones.The long-term seizure outcomes were assessed according to the Engel classification scheme.Results Visual symptoms were present in 25 patients preoperatively in this series.MRI displayed occipital lobe lesions in 15 patients,and FDG-PET revealed hypometabolism in or adjacent to epileptogenic zones.And 30 patients' epileptogenic zones and functional areas were defined by intracranial EEG monitoring.Visual field deficits were present in 35.3% of patients preoperatively,and 61% had new or aggravated visual field deficits after surgery.After a mean follow-up of 41 months,81.1% of the patients were seizure free or rarely had seizures.Conclusion The curative effect of the surgery on the medically intractable occipital lobe epilepsy is good.Intracranial EEG monitoring with electrodes extensively covering the occipital lobe and adjacent areas can be useful to demarcate the epileptogenic zones and the visural cortex,and it may prevent aggravation of the visual field deficits as much as possible.

OBJECTIVETo explore the effects of angiopoietins (Ang-1 and Ang-2) and Tie-2 expression on microvessel density (MVD) in gastric cancers.

METHODSBy using semiquantitative RT-PCR, immunohistochemistry and image analysis system, the expression of Ang-1, Ang-2, Tie-2 mRNA and their proteins were detected in 68 primary gastric cancers and their adjacent normal tissues. Microvessel density (MVD) was figured out based on CD34 immunohistochemical staining.

RESULTSThe expression of all Ang-1, Ang -2, Tie-2 mRNA and their proteins was detected in gastric cancers and their paired adjacent gastric mucosa tissues. A negative correlation between Ang-1 protein, Tie-2 mRNA and MVD in gastric cancers was observed (r = -0.440, r = -0.267; P < 0.05), while the relation between Ang-2 mRNA and its protein, Ang-2/Ang-1 protein ratio with MVD were positive (r = 0.319, r = 0.729, r = 0.739; P < 0.05). It was found that MVD in groups with Ang-2 mRNA T/N ratio over 1.2 (the ratio of Ang-2 mRNA in gastric cancers and its adjacent normal mucosa) was higher than that in those with a ratio under 1.2, revealed by analysing the effects of Ang-1 and Ang-2 mRNA T/N ratio on MVD in gastric cancers.

CONCLUSIONAng-1 activates Tie-2 receptor, whereas Ang-2 antagonizes Ang-1 in the angiogenesis, and the Ang-2/Ang-1 ratio determines angiogenesis and tumor growth in gastric cancers. When the expression of Ang-2 is high and Ang-1 is low, the angiogenesis in gastric cancers is promoted, otherwise oppositely. The role of Ang-2 is dominant in the effect of Angs and their receptor on angiogenesis in gastric cancers.

Angiopoietin-1 ; biosynthesis ; genetics ; Angiopoietin-2 ; biosynthesis ; genetics ; Female ; Humans ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; etiology ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, TIE-2 ; biosynthesis ; genetics ; Stomach Neoplasms ; blood supply ; metabolism ; pathology

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

Adolescent ; Aortic Valve ; abnormalities ; Aortic Valve Insufficiency ; complications ; pathology ; surgery ; Aortic Valve Stenosis ; complications ; pathology ; surgery ; Child ; Endocarditis ; complications ; pathology ; surgery ; Female ; Heart Defects, Congenital ; complications ; pathology ; surgery ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Rheumatic Heart Disease ; complications ; pathology ; surgery

Objective To observe the clinical efficacy of acupuncture plus medicine and function training in treating post-stroke shoulder-hand syndrome. Method Sixty patients with post-stroke shoulder-hand syndrome were randomized into a treatment group of 30 cases and a control group of 30 cases. The treatment group was intervened by acupuncture plus medicine and functional training, while the control group was only by functional training. Before and after intervention, the hand swelling degree, Visual Analogue Scale (VAS), shoulder joint motion scale, Fugl-Meyer Assessment (FMA) scale were used to measure the swelling and pain of hand, motion of shoulder, and motor function of upper limb. Result After intervention, the swelling and pain of hand, motion range of shoulder and motor function of upper limb were significantly improved in both groups (P<0.01), and the improvements in the treatment group were more significant than that in the control group (P<0.05). Conclusion Acupuncture plus medicine and function training can enhance the efficacy in treating post-stroke shoulder-hand syndrome, superior to pure function training.